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. 2009 Feb 27;8(4):586–594. doi: 10.1128/EC.00376-08

FIG. 4.

FIG. 4.

The overproduction of Dse1 in wild-type cells (strain BY4741) increased the level of Slt2 phosphorylation and affected the bud site selection. (A) The cells overproducing Dse1 displayed increased Slt2 phosphorylation in comparison with that of the control cells. Protein extracts were prepared from the wild-type (WT) strain carrying either the empty vector pYC2/CT (strain CRY827) or the pGAL-DSE1-Myc plasmid (overexpressing DSE1 [WT+pGAL-DSE1]; strain CRY826) cultivated overnight in the inducing SCGal-Ura medium. Phosphorylated Slt2 (Phospho-Slt2) was detected by the anti-phospho-p44/42 MAPK antibody (top panel). After stripping, we used the specific anti-Mpk1 antibody to quantify the amount of Slt2 protein (bottom panel). (B) Comparison of the budding patterns of wild-type cells with empty vector (WTa+pGAL; strain CRY827 carrying pYC2/CT) and wild-type cells overexpressing DSE1 (WTa+pGAL-DSE1; strain CRY826 carrying the pGAL-DSE1-Myc plasmid). The wild-type cells overexpressing DSE1 showed the enlarging bud or multiple bud scars within the birth scar. The cells were cultivated overnight in the inducing SCGal-Ura medium and double labeled with calcofluor white and WGA-FITC. The arrows indicate the birth scars. DIC, differential interference contrast. Bar, 5 μm.